PL126606B1 - Method for producing (6r,7r)-7/(z)-2(2-aminothiazol-4-yl)-2-(2-carboxyprop-2-oxyimino)acetamido/-3-(1-pyridinium-methyl)-ceph-3-em-4-carboxylate - Google Patents

Description

The subject of the invention is a method for preparing a new pentahydrate (6R,7R) 1-pyridiniomethyl)-cefemo-3-carboxylate-4 of formula 1. 5 It was found that the compound of formula 1, called ceftazidine or GR 20263, has a wide range of antibiotic activity, and in particular, extremely high activity against Gram-negative microorganisms, including many B-lactamase-producing strains of microorganisms. Gram-negative as described in British Patent No. 2,025,398. The compound has excellent activity against microorganisms that are usually difficult to combat with 6-lactam antibiotics, such as indole-positive Proteus, Serratia, Providence and especially Pseudomonas. This activity is not impaired by human blood serum. Moreover, the reaction of increased inhibition against compound 20 is small and the compound has a rapid bactericidal effect at a concentration close to the lowest inhibitory concentration. The compound distributes well in the bodies of small rodents, producing good therapeutic levels when administered subcutaneously. 25 The compound was successfully used to treat experimental infections with Gram-negative bacteria in mice, obtaining excellent protection against strains of Pseudomonas aeruginosa, which is not particularly sensitive to cephalosporin antibiotics. The level of action was comparable to that of an aminoglycoside such as amikacin. In acute toxicity tests in mice, it turned out that the above compound had an LD^ of more than 6 g/kg. No nephrotoxic effects were observed in rats at doses of 2.0 g/kg. In studies performed on volunteers, the compound showed good pharmacokinetic properties, producing high and long-lasting serum levels after injection. The long half-life in serum suggests the possibility of less frequent administration for less serious infections. Preliminary results of clinical trials show that this compound is characterized by excellent antibiotic properties in vitro and in experimental animals. The British patent description No. 2,025,398 also describes solvates and non-toxic salts, basic and acid addition salts, of the discussed cephalosporins. The British patent specification No. 2,025,398 discloses, among other things, a method for preparing a cephalosporin of formula I and its solvates and non-toxic salts. The method consists in acylation of a compound of formula 2, preferably in the form of a dihydrochloride, with an acid of formula 3, in which R1 is an amino group or a protected amino group, and R2 is a protecting group 126 606126 606 3 4 carboxyl group, or an acylating agent derived from this acid. The protecting group of the amino group and the JEL group can then be removed. The protective carboxyl group, and possibly converting the carboxyl group into a non-toxic salt. A, * The compound of formula 1 according to the British patent description, No. 2*025,398, was obtained in the form of an amorphous substance with insufficient stability, especially at elevated temperature. It was found that /6R,7R/-7-/[/Z The /-2-/2-aminotiazolyl--4/-2-/2-kaifoxypriop-BTOxidimind/iaceftaimido]-3-/1--pyridiniomethyl/-cefemo-3-carboxylate-4 of the formula 1 can be prepared and isolated in the form crystalline pentahydrate, which is the subject of the invention. The new hydrate of the cephalosporin compound of formula 1 is characterized by the highest quality in terms of crystalline form and stability, as well as higher purity. In particular, it was found that the new the hydrate has a clearly defined crystalline structure and is very stable, even when kept for a long period of time at a temperature of 50°C. These properties are valuable in pharmaceutical applications. It was found that the new crystalline hydrate can be easily prepared from solution a salt of the cephalosporin compound in question. Method for preparing the bakehydrate /6R,7R/-7^[/Z/-2-/2-ammothiazole-4/- 30 -2-/2-carboxyprop-2-oxyimino/acetamido]-3-/l-pyridiniomethyl /cephemo-3-carboxylate-4 according to the invention consists in adjusting the pH of the aqueous solution of the free acid or basic salt of the cephalosporin to 2.7-4.8, more precisely 3.0-4.0, preferably 3.3-4.0, e.g. about 3.7, and the desired pentahydrate crystallizes. For example, it has been found that the desired crystalline hydrate can be precipitated from an aqueous solution of an acid addition salt by adding an organic base to obtain a pH value in the above-mentioned range, preferably 3.0-4.0.' ¦ ¦ ' ¦ ) ¦ Precipitation bases that can be used include, for example, inorganic bases such as hydroxides, carbonates or bicarbonates of alkali metals or alkaline earth metals. The starting acid addition salt can be obtained by reacting the cephalosporin with an organic or inorganic acid. Examples of organic acids are carboxylic and sulfonic acids, such as formic, trifluoroacetic, p-toluenesulfonic or methanesulfonic. e.g. use hydrogen chloride, hydrogen bromide, sulfuric acid, nitric acid or phosphoric acid. A particularly suitable acid addition salt is the dihydrochloride of the compound of formula I which can be obtained in very pure form. Alternatively, the new pentahydrate may be prepared by adding an acid to an aqueous solution of the basic salt of the compound of formula I until pH determined in advance, preferably 3.5^4.2. Organic and inorganic acids such as hydrogen chloride or sulfuric acid can be used for this purpose. §5 The starting basic salts may be, for example, alkali metal salts such as sodium or potassium; alkaline earth metal salts such as calcium; salts with amino acids such as lysine or arginine; and salts with organic bases, op. ammonium, methylamine, procaine, phenethylbenzylamine, dibenzylethylenediamine, ethanolairine or N-methylglucosamine. The aqueous medium used may contain a water-miscible organic solvent, in an amount e.g. up to 60% by volume. Examples of organic solvents are ¬ such as alcohols, e.g. ethahol or isopropanol, ethers, e.g. tetrahydrofuran, dioxane or ethyl ether, esters, amides, e.g. N,N-dimethylformamide or N,N-dimethylacetamide, sulfoxides, e.g. dimethylsulfoxide, and nitriles, e.g. acetonitrile. Precipitation is preferably carried out at a temperature of 0-50°C, e.g. 15-40°C, preferably about 24°C. Then, if necessary, the mixture is cooled to a temperature of 0-10°C to increase crystallization yield. After precipitation, the pentahydrate is filtered off, washed and dried using conventional methods. For example, the hydrate may be air dried, dried under reduced pressure, or preferably dried in a sterile inert gas such as sterile nitrogen. Care must be taken during drying so that the water content of the product does not fall too low. The salts for use in the preparation of the new pentahydrate according to the invention can be obtained in the manner described in the British patent specification No. 2,025,398, i.e. corresponding to that described above. The hydrochloride can be obtained in the form of a very stable crystalline dichloride by crystallization from a medium containing acetone and formic acid, i.e. under the conditions described in Example 1. The new crystalline pentahydrochloride according to the invention was subjected to X-ray analysis. The product from Example 4 was used to prepare a powder diagram using the Debye and Scherre method, exposing it to CoKa radiation for 12 hours. The second diagram was made by exposing the sample to CuKa radiation for 3 hours. The line intensities were compared with a set of standards and the obtained values of relative intensities are listed in Table 1. The new pentahydrate according to the invention was also characterized by an infrared spectrum. The infrared spectrum in the paraffin oil of the product from Example 4 is shown in the attached drawing. The crystalline pentahydrate according to the invention has the antibiotic properties of a compound of formula 1 and can be used to combat various diseases caused by pathogenic bacteria in humans and animals, such as respiratory diseases and urinary tract diseases. The invention also relates to pharmaceutical compositions containing the new pentahydrate, with ¬ used for use in human medicine126 606 5 6 Table 1 ddO-inm) 15.9 9.9 ¦ 8.7 7.9 6.7 6.3 | 5.95 5.74 5.42 ' -5.18 4.71 \ 4.50 1 4.37 4.15 1 4.10 1 4.01 3.93 3.86 3.68 3.41 3, 33 3.29 Intensity: medium high, high trace, medium diffuse, weak, medium weak, weak, medium, medium, trace, large, medium, weak, large, weak, very slight, b. slight ddO-inm) 3.24 3.18 3, 13 3.01 2.87 2.77 2.72 2.69 2.52 2.43 2.33 2.29 2.26 2.23 2.19 27lT 2.07 2.03 1.97 1.94 Intensity very slight weak weak medium medium medium weak weak diffused medium weak weak very weak very weak very weak weak very weak very weak very weak very weak v.___ and animal. Such compositions can be prepared in the usual manner using the necessary pharmaceutical carriers and diluents. The antibiotic pentahydrate according to the invention can be prepared for injection in unit doses, in ampoules or in containers for repeated use, optionally with the addition of preservatives. Others can also be prepared preparations such as suspensions, solutions or suspensions in oily or aqueous vehicles. These preparations may contain various additional substances, such as suspending, stabilizing and/or dispersing agents. It is desirable that the active ingredient be in the form of a powder which can be mixed with a suitable vehicle, e.g. sterile and pyrogenic water. Advantageously, the preparation may contain a suitable non-toxic base to improve the water solubility of the active ingredient and /or ensuring that it remains in an aqueous solution at a physiologically acceptable pH value. Alternatively, the base may be present in the water in which the powder is dissolved. As bases, for example, inorganic bases such as sodium carbonate, sodium bicarbonate, trisodium orthophosphate or sodium sulfite, or organic bases such as lysine, lysine acetate, tromethamine, arginine or sodium glycinate may be used. preparations in the form of suppositories, e.g. usually containing suppository bases such as cocoa butter or other glycerides. For use in the eyes or ears, individual capsules may be prepared in liquid or semi-liquid form or as drops. The compositions may contain from 0, 1%, e.g. 0.1-99% of the active ingredient, depending on the method of administration. In the case of unit doses, each of them should preferably contain 50-1500 mg of the active ingredient. The dose for adults is 1000-3000 mg/day. To combat infections depending on the route and frequency of administration. For example, in adult patients, a sufficient dose when administered intravenously or intramuscularly is 1000-3000 mg/day. A higher daily dose may be required to combat infections caused by Pseudomonas. The antibiotic of the invention may be administered in combination with other therapeutic agents such as antibiotics, e.g. penicillins or other cephalosporins. The examples given below serve to illustrate the method of preparation. pentahydrate according to the invention. Examples 1 and 2 describe the preparation of the starting acid addition salts for the production of the pentahydrate. In all examples, the abbreviation TFA stands for trifluoroacetic acid. Water content was determined using the Karl Fischer method. CSF spectra were taken at 100 MHz. J values are given in Hz and the abbreviations mean respectively: s - singlet, t - triplet, d - doublet, dd - doublet of doublets, m - multiplet, ABq - quartet AB. Amberlite L.A.2 is a weakly basic high molecular weight secondary amine produced by Rohm and Haas, Philadelphia, St. American food Hyflo Super-Cel is a diatomaceous earth used as a filtration aid, manufactured by Johms-Mainyille, St. I'll eat it. Am. Example I. Preparation of dihydrochloride (6R,7R)-7-[/Z/-2-/2-aminotiazolyl-4]-2-/2-carboxy-prop-2-oxyimino/-acetamido]-3-/ 1-pyridiniomethyl-/cefemo-3-carboxylate-4. Up to 41.8 g of N,N-dimethylformamide solvate (6R,7R)-7-[2-/2-tritylaminotiazolyl-4/-2-/2-III- n-butoxycarbonylprop-2-oxyimino/acetamido]-3-/1-pyridiniomethyl/cephemo-3-carboxylate-4 while stirring and cooling, 84 ml of formic acid were added to the water to maintain the temperature below 28°C. The obtained solution cooled to 20°C and 17 ml of concentrated hydrochloric acid were added dropwise within 5 minutes. The mixture was stirred for 3 hours at room temperature, and then the precipitated triphenylmethanol was filtered off. The filtrate was added with stirring to 800 ml of acetone. The triphenylmethanol was washed with 3 x 7 ml of formic acid and the filtrate was added to a mixture of the previous filtrate and acetone. The resulting suspension was stirred for 1.25 hours and then filtered. The crystalline precipitate was washed with acetone and dried under reduced pressure to give 20.2 g of the title compound. Chlorine content: calculated for C22H24N6CtS2- C1&:. ll^/o, found: 11.0%, \Ultraviolet spectrum: ?*ax (pH G, phosphate buffer) 257 nm (Ej* cra = 347), X inflection ^teni (E^cm = 310) and 290 nm (Ej* cm = = 150).- ¦Pxzy clade II; Hydrogen sulphate (6R,7R)-7^[/Z-2- */2-aminotiazolylOT4/-2-/2-carboxyprop-2-oxyimino/acetamido]-3-/l-pyridiniomethyl/cefemo-3-carbo - xylan-4. a) Up to 80 g of N,N-dimethylformamide solvate (6R,7R) (acetami-cto]-3-(1-pyridiniomethyl)cephemo-3-carboxylate-4 150 ml of 98^100% formic acid were added. The mixture was stirred until a clear solution was obtained, then cooled to 15°C and 12.6 ml of water and 12.6 ml of concentrated sulfuric acid were added. The temperature rose to 25°C. Stirring was continued at this temperature for 5 hours. The suspension was filtered and the precipitate was washed with 46 ml of 98-100% formic acid in several portions. The Felaezone filtrate was diluted with 200 ml of isopropanol and the obtained solution was added dropwise over 20 minutes to 1400 ml of isopropanol. 400 ml of isopropyl ether were added to the obtained suspension and the mixture was stirred for 10 minutes, then the precipitate was filtered off, washed with 3 x 200 ml of isopropanol and dried for 16 hours under reduced pressure at 40°C. 51.02 g of the title compound were obtained in the form of a cream-colored precipitate. The water content determined by the Karl Fischer method was 4.2%, calculated as 5.3%. The sulfur content is 15.2%, calculated for C22H22N607S2H2S04 • • 2HtO - 14.13%. b) The product from step a) was gently stirred and heated in 10 ml of methanol, after which the obtained suspension was left for 2 hours at room temperature and then for 30 minutes at 0°C. The precipitate was filtered off, washed with cold methanol and dried for 3 hours under reduced pressure at 40°C. 0.9 g of the crystalline title compound were obtained. Elemental analysis: calculated for C22H2zN607S2H2S04 • 2H20 : C - 38.8 H — 4.15, N — 12.34, S — 14.13, H-jO — 5.3; found: C - 38.53, H - 3.70, N - 12.36, S - 14.1, H20 - 4.5. Ultraviolet spectrum: Xmax (pH6, phosphate buffer) 257 nm (E^cm = 325), 241 nm (e\%cm = 292), 289 nm (E}* cm = 144), CSF spectrum/TFA/t: 0.90/d, J = = 6 Hz, 2H), 1.32 /t, J = 7 Hz, lH/i 1.80/t, J = 7 Hz, 2H) (pyridyl protons), 1.40/d, J = 8 Hz, 1H, NHCO), 2.60/s , 1H, thiazolyl proton), 3.72 and 4.50 (ABq, J = 14 Hz, 2H, group -CHjN =), 3.84/m, 1H, 7-H), 4.54 (d, J = 5Hz), 6-H), 6.06 and 6.54 (ABq, J=il8 Hz, 2H, 2-H), and 8.20 (s, 6H, C/CH,/2). All following the examples illustrate the preparation of the pentahydrate /6Rr7R/-7-^Z/-2-/2-aminotiazolyl-4/-2-/2-kajr]x)jksyipT PL PL PL PL PL PL

Claims (4)

1. Patent claims 1. Method for preparing the new pentahydrate /6R,7R/-7-<[/Z/-2-/2-amiiniothiazolyl-4/2-/2-kairbo(ksy prop-2-oksy(imdTio/acetaimiido) ]-3-/1-pyrid(niomethyl/cefemo-3-carboxylate-4), characterized in that the pH of the solution of the free acid or basic salt /6R,7R/-7-[/Z/-2- (2-ami'notiazolyl-4/2-/2Hcarboxyprop-2-oxyimino/acetamido]-3-/1-pyridinomethyl/cefemo-3-carboxylate-4 in an aqueous medium- Table 3 pH 3.2 | 3.4 | 3.8 | 4.2 4.7 4.8 Yield (% mol) 18.8 52.8 82.8 72.0 21.0 8.8 Water content in % 13.6 13.65 13.35 13.95 13.7 is adjusted to 2.7-4.8 and the given pentahydrate crystallizes. 2. The method according to claim 1, characterized in that the acid addition salt (6R,7R) -7-[/Z/-2-/2-ammothiazolflo-4/-2-/2-carboxyprop-2-oxyimino//acetaimido]^ 3-(1-py3ndyiniimethyl)pheno-3-carboxylic acid and the pH is adjusted to 3.0-4.0. 3. The method according to claim 1, characterized in that the basic salt (6R,7R)-7-[/Z/-2-/2-aminotiazoine/-4-2-/2-kaiboxyprop-;2r<^xytoine/ace! taim!i- to]-3-(1-pyridylmethyl)cephemo-3-carboxylate-4 and the pH is adjusted to 3.5-4.2. 4. The method according to claim 1, characterized in that the pH of the solution of the free acid or base salt of [-Acetaimido]-3-(1-pyridiniomethyl)cefemo-3-carboxylaou-4 in an aqueous medium is adjusted to 3.3-4.0 and the given pentahydrate is crystallized. 10 15 20 25 i126 606 NH, S ' • N .CC ti ^ O estrus. I 3 xO-C-COOL I CH3 PATTERN 1 • S- N -CH2"0N^ coo H H H9N—r-f -N C cooG R COOH CH 3 N C.COOR C 2 3 PATTERN 3 Fig. 1 ZGK 5 Btm, order 9124 - 80 copies. Price PLN 100 PL PL PL PL PL PL PL PL